REGULATION OF APICAL MEMBRANE CL CHANNELS IN AIRWAYS

  • Widdicombe, Jonathan (PI)
  • Wine, Jeffrey (PI)
  • Szoka, Francis (PI)
  • Verkman, Allan (PI)
  • Finkbeiner, Walter (PI)

Project: Research project

Project Details

Description

The research proposed here concentrates on the regulation of apical
membrane C1 channels in airways. In addition a novel genetic
approach to the localization of the defective gene on chromosome
7 will take advantage of the screening methods used in the ion
transport studies. Cultured cells will be used throughout this
proposal. In Project 1 (Widdicombe), levels of second messengers,
protein kinases and their target phosphoproteins in normal and CF
cells will be compared. Changes in protein phosphorylation will
be compared with changes in C1 secretion. In Project 2 (Wine),
regulation of C1 channels will be studied using a variety of
techniques including patch-clamping. Emphasis will be placed on
determining if the same C1 channel defect found in airways can be
demonstrated in other affected epithelia, and on whether more than
one channel type is affected. In Project 3 (Verkman), regulation
of the C1 channel will be studied following reconstitution into
liposomes or planar lipid bilayers. Apical membrane vesicles from
airway cultures and other tissues will be used as sources of C1
channels. Attempts will be made to purify the C1 channel, though
this is not necessary for successful reconstitution. In
reconstitution studies, the C1 channel is effectively separated
from other apical membrane proteins, providing a direct means of
testing whether it is defective in CF. Clinical research will be
performed at a clinical/cell acquisition CORE in stanford and human
cell culture CORE at each university. The clinical/cell
acquisition CORE (Lewiston) will provide airway tissues for the
USCF Culture CORE (Finkbeiner). Attempts will be made to transform
the cells and to improve the level of differentiation of the
cultures as revealed by electrophysiological studies. Ultimately,
the clinical/cell acquisition CORE will apply information obtained
in the basic science projects to the treatment of patients with CF.
StatusFinished
Effective start/end date9/30/888/31/99

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

ASJC

  • Medicine(all)

Fingerprint Explore the research topics touched on by this project. These labels are generated based on the underlying awards/grants. Together they form a unique fingerprint.